Rastering strategy for screening and centring of microcrystal samples of human membrane proteins with a sub-10 µm size X-ray synchrotron beam by Vadim.

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Presentation transcript:

Rastering strategy for screening and centring of microcrystal samples of human membrane proteins with a sub-10 µm size X-ray synchrotron beam by Vadim Cherezov, Michael A. Hanson, Mark T. Griffith, Mark C. Hilgart, Ruslan Sanishvili, Venugopalan Nagarajan, Sergey Stepanov, Robert F. Fischetti, Peter Kuhn, and Raymond C. Stevens Interface Volume 6(Suppl 5):S587-S597 October 6, 2009 ©2009 by The Royal Society

Improvements in size and shape of β2AR-T4L crystals as the result of optimizations of crystallization conditions. Vadim Cherezov et al. J. R. Soc. Interface 2009;6:S587-S597 ©2009 by The Royal Society

Imaging of small A2AR-T4L crystals in LCP. (a) Image recorded with a Zeiss AxioImager microscope using a 10× objective and a bright-field illumination. Vadim Cherezov et al. J. R. Soc. Interface 2009;6:S587-S597 ©2009 by The Royal Society

Imaging of small β2AR-T4L crystals in LCP. (a) Image recorded with RockImager 1000 (Formulatrix) using a bright-field illumination. Vadim Cherezov et al. J. R. Soc. Interface 2009;6:S587-S597 ©2009 by The Royal Society

Images of loops with harvested β2AR-T4L crystals under a cryo-stream recorded through the inline optics at the GM/CA beamline at APS. (a) A nylon loop with several optically invisible β2AR-T4L crystals embedded into a frozen lipidic mesophase. Vadim Cherezov et al. J. R. Soc. Interface 2009;6:S587-S597 ©2009 by The Royal Society

A screenshot of the automated rastering interface window in Blu-Ice. Vadim Cherezov et al. J. R. Soc. Interface 2009;6:S587-S597 ©2009 by The Royal Society

Effect of radiation damage on the intensity of diffraction spots collected in different resolution shells (identified as different colours) from a crystal of A2AR-T4L. Vadim Cherezov et al. J. R. Soc. Interface 2009;6:S587-S597 ©2009 by The Royal Society

Crystal structures of (a) β2AR-T4L bound to carazolol (PDB ID 2RH1; Cherezov et al. 2007), (b) β2AR(E122W)-T4L bound to timolol (PDB ID 3D4S; Hanson et al. 2008) and (c) A2AR-T4L bound to ZM (PDB ID 3EML; Jaakola et al. 2008). β2AR-T4L/carazolol cryst... Vadim Cherezov et al. J. R. Soc. Interface 2009;6:S587-S597 ©2009 by The Royal Society